Skip to Content
Merck
  • MiR-34a targeting of Notch ligand delta-like 1 impairs CD15+/CD133+ tumor-propagating cells and supports neural differentiation in medulloblastoma.

MiR-34a targeting of Notch ligand delta-like 1 impairs CD15+/CD133+ tumor-propagating cells and supports neural differentiation in medulloblastoma.

PloS one (2011-09-21)
Pasqualino de Antonellis, Chiara Medaglia, Emilio Cusanelli, Immacolata Andolfo, Lucia Liguori, Gennaro De Vita, Marianeve Carotenuto, Annamaria Bello, Fabio Formiggini, Aldo Galeone, Giuseppe De Rosa, Antonella Virgilio, Immacolata Scognamiglio, Manuela Sciro, Giuseppe Basso, Johannes H Schulte, Giuseppe Cinalli, Achille Iolascon, Massimo Zollo
ABSTRACT

Through negative regulation of gene expression, microRNAs (miRNAs) can function as oncosuppressors in cancers, and can themselves show altered expression in various tumor types. Here, we have investigated medulloblastoma tumors (MBs), which arise from an early impairment of developmental processes in the cerebellum, where Notch signaling is involved in many of the cell-fate-determining stages. Notch regulates a subset of MB cells that have stem-cell-like properties and can promote tumor growth. On the basis of this evidence, we hypothesized that miRNAs targeting the Notch pathway can regulate these phenomena, and can be used in anti-cancer therapies. In a screening of potential targets within Notch signaling, miR-34a was seen to be a regulator of the Notch pathway through its targeting of Notch ligand Delta-like 1 (Dll1). Down-regulation of Dll1 expression by miR-34a negatively regulates cell proliferation, and induces apoptosis and neural differentiation in MB cells. Using an inducible tetracycline on-off model of miR-34a expression, we show that in Daoy MB cells, Dll1 is the first target that is regulated in MB, as compared to the other targets analyzed here: Cyclin D1, cMyc and CDK4. MiR-34a expression negatively affects CD133(+)/CD15(+) tumor-propagating cells, then we assay through reverse-phase proteomic arrays, Akt and Stat3 signaling hypo-phosphorylation. Adenoviruses carrying the precursor miR-34a induce neurogenesis of tumor spheres derived from a genetic animal model of MB (Patch1(+/-) p53(-/-)), thus providing further evidence that the miR-34a/Dll1 axis controls both autonomous and non autonomous signaling of Notch. In vivo, miR-34a overexpression carried by adenoviruses reduces tumor burden in cerebellum xenografts of athymic mice, thus demonstrating an anti-tumorigenic role of miR-34a in vivo. Despite advances in our understanding of the pathogenesis of MB, one-third of patients with MB remain incurable. Here, we show that stable nucleic-acid-lipid particles carrying mature miR-34a can target Dll1 in vitro and show equal effects to those of adenovirus miR-34a cell infection. Thus, this technology forms the basis for their therapeutic use for the delivery of miR-34a in brain-tumor treatment, with no signs of toxicity described to date in non-human primate trials.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
StableCell MEM, With Earle′s salts, stable glutamine, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
hEGF, EGF, recombinant, expressed in E. coli, lyophilized powder, suitable for cell culture
Sigma-Aldrich
Epidermal Growth Factor from mouse, EGF
Sigma-Aldrich
StableCell MEM, Alpha Modification, With stable glutamine and sodium bicarbonate, with ribonucleosides and deoxyribonucleosides, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
StableCell MEM, Alpha Modification, With stable glutamine and sodium bicarbonate, without ribonucleosides and deoxyribonucleosides, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts, L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Alpha Modification, with sodium bicarbonate, without L-glutamine, ribonucleosides and deoxyribonucleosides, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, 10 ×, With Earle′s salts, without L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts, L-glutamine, and non-essential amino acids, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Modified, with Earle′s salts and reduced NaHCO3. without L-glutamine, liquid, sterile-filtered, suitable for cell culture
SAFC
Minimum Essential Medium Eagle, HEPES Modification, with Earle′s salts, L-glutamine and 25 mM HEPES, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Joklik Modification, with L-glutamine, without calcium chloride and sodium bicarbonate, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Spinner Modification, with Earle′s salts and sodium bicarbonate, without calcium chloride and L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Alpha Modification, with ribonucleosides, deoxyribonucleosides and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Alpha Modification, with L-glutamine and sodium pyruvate, without ribonucleosides, deoxyribonucleosides and sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts and L-glutamine, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, HEPES Modification, with Earle′s salts, 25 mM HEPES and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, 10 ×, With Hanks′ salts, without L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Hanks′ salts and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Alpha Modification, with L-glutamine, ribonucleosides and deoxyribonucleosides, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Hanks′ salts, L-glutamine and non-essential amino acids, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts, non-essential amino acids and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Joklik Modification, With sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, with Earle′s salts and non-essential amino acids, without L-glutamine, phenol red and sodium bicarbonate, Modified, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Hanks′ salts, L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Hanks′ salts and L-glutamine, without sodium bicarbonate, powder, suitable for cell culture
SAFC
Minimum Essential Medium Eagle, AutoMod, with Earle′s salts, without L-glutamine and sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Alpha Modification, With sodium bicarbonate and L-glutamine, without ribonucleosides and deoxyribonucleosides, liquid, sterile-filtered, suitable for cell culture